Tumor necrosis related receptor, TR7

ABSTRACT

TR7 polypeptides and polynucleotides and methods for producing such polypeptides by recombinant techniques are disclosed. Also disclosed are methods for utilizing TR7 polypeptides and polynucleotides in the design of protocols for the treatment of chronic and acute inflammation, arthritis, septicemia, autoimmune diseases (such as inflammatory bowel disease and psoriasis), transplant rejection, graft versus host disease, infection, stroke, ischemia, acute respiratory disease syndrome, restenosis, brain injury, AIDS, bone diseases, cancer (such as lymphoproliferative disorders), atherosclerosis, and Alzheimers disease, among others and diagnostic assays for such conditions.

This application is a division of U.S. application Ser. No. 08/959,382,filed Oct. 28, 1997, U.S. Pat. No. 6,013,476 which claims the benefit ofU.S. Provisional Application Ser. No. 60/041,796, filed Apr. 2, 1997,both of whose contents are incorporated herein by reference in theirentireties.

FIELD OF INVENTION

This invention relates to newly identified polynucleotides, polypeptidesencoded by them and to the use of such polynucleotides and polypeptides,and to their production. More particularly, the polynucleotides andpolypeptides of the present invention relate to Tumor necrosis factorreceptor (TNF-R) family, hereinafter referred to as TR7. The inventionalso relates to inhibiting or activating the action of suchpolynucleotides and polypeptides.

BACKGROUND OF THE INVENTION

Many biological actions, such as responses to certain stimuli andnatural biological processes, are controlled by factors such ascytokines. Cytokines generally act through receptors by engaging thereceptor and producing an intracellular response.

For example, tumor necrosis factors (TNF) alpha and beta are cytokineswhich act through TNF receptors to regulate numerous biologicalprocesses, including protection against infection and induction of shockand inflammatory disease. The TNF molecules belong to the “TNF-ligand”superfamily, and act together with their receptors or counter-ligands,the “TNF-receptor” superfamily. So far, nine members of the TNF-ligandsuperfamily have been identified and ten members of the TNF-receptorsuperfamily have been characterized. Identified ligands include TNF-α,lymphotoxin-α (LT-α, also known as TNF-β), LT-β (found in complexheterotrimer LT-α2-β), FasL, CD40L, CD27L, CD30L, 4-1BBL, OX40L andnerve growth factor (NGF). The superfamily of TNF-receptors include thep55TNF receptor, p75TNF receptor, TNF receptor-related protein, FASantigen or APO-1, CD40, CD27, CD30, 4-1BB, OX40, low a p75 andNGF-receptor (Meager, A., Biologicals, 22:291-295 (1994)).

Many members of the TNF-ligand superfamily are expressed by activatedT-cells, which implies that they are necessary for T-cell interactionswith other cell types which underlie cell ontogeny and functions.Considerable insight into the essential functions of several members ofthe TNF receptor family has been gained from the identification andcreation of mutants which abolish the expression of these proteins. Forexample, naturally occurring mutations in the FAS antigen and its ligandcause lymphoproliferative disease (Watanabe-Fukunaga, R., et al., Nature356:314 (1992)), perhaps reflecting a failure of programmed cell death.Mutations of the CD40 ligand cause an X-linked immunodeficiency statecharacterized by high levels of immunoglobulin M and low levels ofimmunoglobulin G in plasma, indicating faulty T-cell-dependent B-cellactivation (Allen, R. C. et al., Science 259:990 (1993)). Targetedmutations of the low affinity nerve growth factor receptor cause adisorder characterized by faulty sensory innovation of peripheralstructures (Lee, K. F. et al, Cell 69:737 (1992)).

TNF and LT-α are capable of binding to two TNF receptors (the 55- and75-kd TNF receptors). A large number of biological effects elicited byTNF and LT-α acting through their receptors include hemorrhagic necrosisof transplanted tumors, cytotoxicity, endotoxic shock, inflammation,immunoregulation, proliferation and anti-viral responses, as well asprotection against the deleterious effects of ionizing radiation. TNFand LT-α are involved in the pathogenesis of a wide range of diseases,including endotoxic shock, cerebral malaria, tumors, autoimmuinedisease, AIDS and graft-versus-host rejection (Beutler, B. and VonHuffel, C., Science 264:667-668 (1994)). Mutations in the p55 Receptorcause increased susceptibility to microbial infection. Moreover, anapproximately 80 amino acid domain near the C-terminus of TNFR1 (P55)and Fas has been reported as the “death domain,” which is responsiblefor transducing signals for programmed cell death (Tartaglia et al.,Cell 74:845 (1993)).

The effects of TNF family ligands and TNF family receptors are variedand influence numerous functions, both normal and abnormal, in thebiological processes of the mammalian system. There is a clear need,therefore, for identification and characterization of such receptors andligands that influence biological activity, both normally and in diseasestates. In particular, there is a need to isolate and characterize novelmembers of the TNF receptor family.

This indicates that these receptors have an established, proven historyas therapeutic targets. Clearly there is a need for identification andcharacterization of further receptors which can play a role inpreventing, ameliorating or correcting dysfunctions or diseases,including, but not limited to, chronic and acute inflammation,arthritis, septicemia, autoimmune diseases (such as inflammatory boweldisease and psoriasis), transplant rejection, graft versus host disease,infection, stroke, ischemia, acute respiratory disease syndrome,restenosis, brain injury, AIDS, bone diseases, cancer (such aslymphoproliferative disorders), atherosclerosis, and Alzheimers disease.

SUMMARY OF THE INVENTION

In one aspect, the invention relates to TR7 polypeptides and recombinantmaterials and methods for their production. Another aspect of theinvention relates to methods for using such TR7 polypeptides andpolynucleotides. Such uses include the treatment of chronic and acuteinflammation, arthritis, septicemia, autoimmune diseases (such asinflammatory bowel disease and psoriasis), transplant rejection, graftversus host disease infection, stroke, ischemia, acute respiratorydisease syndrome, restenosis, brain injury, AIDS, bone diseases, cancer(such as lymphoproliferative disorders), atherosclerosis, and Alzheimersdisease among others. In still another aspect, the invention relates tomethods to identify agonists and antagonists using the materialsprovided by the invention, and treating conditions associated with TR7imbalance with the identified compounds. Yet another aspect of theinvention relates to diagnostic assays for detecting diseases associatedwith inappropriate TR7 activity or levels.

DESCRIPTION OF THE INVENTION

Definitions

The following definitions are provided to facilitate understanding ofcertain terms used frequently herein.

“TR7” refers, among others, to a polypeptide comprising the amino acidsequence set forth in SEQ ID NO:2, or an allelic variant thereof.

“Receptor Activity” or “Biological Activity of the Receptor” refers tothe metabolic or physiologic function of said TR7 including similaractivities or improved activities or these activities with decreasedundesirable side-effects. Also included are antigenic and immunogenicactivities of said TR7.

“TR7 gene” refers to a polynucleotide comprising the nucleotide sequenceset forth in SEQ ID NO:1 or allelic variants thereof and/or theircomplements.

“Antibodies” as used herein includes polyclonal and monoclonalantibodies, chimeric, single chain, and humanized antibodies, as well asFab fragments, including the products of an Fab or other immunoglobulinexpression library.

“Isolated” means altered “by the hand of man” from the natural state. Ifan “isolated” composition or substance occurs in nature, it has beenchanged or removed from its original environment, or both. For example,a polynucleotide or a polypeptide naturally present in a living animalis not “isolated,” but the same polynucleotide or polypeptide separatedfrom the coexisting materials of its natural state is “isolated”, as theterm is employed herein.

“Polynucleotide” generally refers to any polyribonucleotide orpolydeoxribonucleotide, which may be unmodified RNA or DNA or modifiedRNA or DNA. “Polynucleotides” include, without limitation single- anddouble-stranded DNA, DNA that is a mixture of single- anddouble-stranded regions, single- and double-stranded RNA, and RNA thatis mixture of single- and double-stranded regions, hybrid moleculescomprising DNA and RNA that may be single-stranded or, more typically,double-stranded or a mixture of single- and double-stranded regions. Inaddition, “polynucleotide” refers to triple-stranded regions comprisingRNA or DNA or both RNA and DNA. The term polynucleotide also includesDNAs or RNAs containing one or more modified bases and DNAs or RNAs withbackbones modified for stability or for other reasons. “Modified” basesinclude, for example, tritylated bases and unusual bases such asinosine. A variety of modifications has been made to DNA and RNA; thus,“polynucleotide” embraces chemically, enzymatically or metabolicallymodified forms of polynucleotides as typically found in nature, as wellas the chemical forms of DNA and RNA characteristic of viruses andcells. “Polynucleotide” also embraces relatively short polynucleotides,often referred to as oligonucleotides.

“Polypeptide” refers to any peptide or protein comprising two or moreamino acids joined to each other by peptide bonds or modified peptidebonds, i.e., peptide isosteres. “Polypeptide” refers to both shortchains, commonly referred to as peptides, oligopeptides or oligomers,and to longer chains, generally referred to as proteins. Polypeptidesmay contain amino acids other than the 20 gene-encoded amino acids.“Polypeptides” include amino acid sequences modified either by naturalprocesses, such as posttranslational processing, or by chemicalmodification techniques which are well known in the art. Suchmodifications are well described in basic texts and in more detailedmonographs, as well as in a voluminous research literature.Modifications can occur anywhere in a polypeptide, including the peptidebackbone, the amino acid side-chains and the amino or carboxyl termini.It will be appreciated that the same type of modification may be presentin the same or varying degrees at several sites in a given polypeptide.Also, a given polypeptide may contain many types of modifications.Polypeptides may be branched as a result of ubiquitination, and they maybe cyclic, with or without branching. Cyclic, branched and branchedcyclic polypeptides may result from posttranslation natural processes ormay be made by synthetic methods. Modifications include acetylation,acylation, ADP-ribosylation, amidation, covalent attachment of flavin,covalent attachment of a heme moiety, covalent attachment of anucleotide or nucleotide derivative, covalent attachment of a lipid orlipid derivative, covalent attachment of phosphotidylinositol,cross-linking, cyclization, disulfide bond formation, demethylation,formation of covalent cross-links, formation of cystine, formation ofpyroglutamate, formylation, gamma-carboxylation, glycosylation, GPIanchor formation, hydroxylation, iodination, methylation,myristoylation, oxidation, proteolytic processing, phosphorylation,prenylation, racemization, selenoylation, sulfation, transfer-RNAmediated addition of amino acids to proteins such as arginylation, andubiquitination. See, for instance, PROTEINS—STRUCTURE AND MOLECULARPROPERTIES, 2nd Ed., T. E. Creighton, W. H. Freeman and Company, NewYork, 1993 and Wold, F., Posttranslational Protein Modifications:Perspectives and Prospects, pgs. 1-12 in POSTTRANSLATIONAL COVALENTMODIFICATION OF PROTEINS, B. C. Johnson, Ed., Academic Press, New York,1983; Seifter et al., “Analysis for protein modifications and nonproteincofactors”, Meth Enzymol (1990) 182:626-646 and Rattan et al., “ProteinSynthesis: Posttranslational Modifications and Aging”, Ann NY Acad Sci(1992) 663:48-62.

“Variant” as the term is used herein, is a polynucleotide or polypeptidethat differs from a reference polynucleotide or polypeptiderespectively, but retains essential properties. A typical variant of apolynucleotide differs in nucleotide sequence from another, referencepolynucleotide. Changes in the nucleotide sequence of the variant may ormay not alter the amino acid sequence of a polypeptide encoded by thereference polynucleotide. Nucleotide changes may result in amino acidsubstitutions, additions, deletions, fusions and truncations in thepolypeptide encoded by the reference sequence, as discussed below. Atypical variant of a polypeptide differs in amino acid sequence fromanother, reference polypeptide. Generally, differences are limited sothat the sequences of the reference polypeptide and the variant areclosely similar overall and, in many regions, identical. A variant andreference polypeptide may differ in amino acid sequence by one or moresubstitutions, additions, deletions in any combination. A substituted orinserted amino acid residue may or may not be one encoded by the geneticcode. A variant of a polynucleotide or polypeptide may be a naturallyoccurring such as an allelic variant, or it may be a variant that is notknown to occur naturally. Non-naturally occurring variants ofpolynucleotides and polypeptides may be made by mutagenesis techniquesor by direct synthesis.

“Identity” is a measure of the identity of nucleotide sequences or aminoacid sequences. In general, the sequences are aligned so that thehighest order match is obtained. “Identity” per se has an art-recognizedmeaning and can be calculated using published techniques. See, e.g.:(COMPUTATIONAL MOLECULAR BIOLOGY, Lesk, A. M., ed., Oxford UniversityPress, New York, 1988; BIOCOMPUTING: INFORMATICS AND GENOME PROJECTS,Smith, D. W., ed., Academic Press, New York, 1993; COMPUTER ANALYSIS OFSEQUENCE DATA, PART I, Griffin, A. M., and Griffin, H. G., eds., HumanaPress, New Jersey, 1994; SEQUENCE ANALYSIS IN MOLECULAR BIOLOGY, vonHeinje, G., Academic Press, 1987; and SEQUENCE ANALYSIS PRIMER,Gribskov, M. and Devereux, J., eds., M Stockton Press, New York, 1991).While there exist a number of methods to measure identity between twopolynucleotide or polypeptide sequences, the term “identity” is wellknown to skilled artisans (Carillo, H., and Lipton, D., SIAM J AppliedMath (1988) 48:1073). Methods commonly employed to determine identity orsimilarity between two sequences include, but are not limited to, thosedisclosed in Guide to Huge Computers, Martin J. Bishop, ed., AcademicPress, San Diego, 1994, and Carillo, H., and Lipton, D., SIAM J AppliedMath (1988) 48:1073. Methods to determine identity and similarity arecodified in computer programs. Preferred computer program methods todetermine identity and similarity between two sequences include, but arenot limited to, GCS program package (Devereux, J., et al., Nucleic AcidsResearch (1984) 12(1):387), BLASTP, BLASTN, FASTA (Atschul, S. F. etal., J Molec Biol (1990) 215:403).

As an illustration, by a polynucleotide having a nucleotide sequencehaving at least, for example, 95% “identity” to a reference nucleotidesequence of SEQ ID NO: 1 is intended that the nucleotide sequence of thepolynucleotide is identical to the reference sequence except that thepolynucleotide sequence may include up to five point mutations per each100 nucleotides of the reference nucleotide sequence of SEQ ID NO: 1. Inother words, to obtain a polynucleotide having a nucleotide sequence atleast 95% identical to a reference nucleotide sequence, up to 5% of thenucleotides in the reference sequence may be deleted or substituted withanother nucleotide, or a number of nucleotides up to 5% of the totalnucleotides in the reference sequence may be inserted into the referencesequence. These mutations of the reference sequence may occur at the 5or 3 terminal positions of the reference nucleotide sequence or anywherebetween those terminal positions, interspersed either individually amongnucleotides in the reference sequence or in one or more contiguousgroups within the reference sequence.

Similarly, by a polypeptide having an amino acid sequence having atleast, for example, 95% “identity” to a reference amino acid sequence ofSEQ ID NO:2 is intended that the amino acid sequence of the polypeptideis identical to the reference sequence except that the polypeptidesequence may include up to five amino acid alterations per each 100amino acids of the reference amino acid of SEQ ID NO: 2. In other words,to obtain a polypeptide having an amino acid sequence at least 95%identical to a reference amino acid sequence, up to 5% of the amino acidresidues in the reference sequence may be deleted or substituted withanother amino acid, or a number of amino acids up to 5% of the totalamino acid residues in the reference sequence may be inserted into thereference sequence. These alterations of the reference sequence mayoccur at the amino or carboxy terminal positions of the reference aminoacid sequence or anywhere between those terminal positions, interspersedeither individually among residues in the reference sequence or in oneor more contiguous groups within the reference sequence.

Polypeptides of the Invention

In one aspect, the present invention relates to TR7 polypeptides (or TR7proteins). The TR7 polypeptides include the polypeptides of SEQ ID NOS:2and 4; as well as polypeptides comprising the amino acid sequence of SEQID NO:2; and polypeptides comprising the amino acid sequence which haveat least 80% identity to that of SEQ ID NO:2 over its entire length, andstill more preferably at least 90% identity, and even still morepreferably at least 95% identity to SEQ ID NO: 2. Furthermore, thosewith at least 97-99% are highly preferred. Also included within TR7polypeptides are polypeptides having the amino acid sequence which haveat least 80% identity to the polypeptide having the amino acid sequenceof SEQ ID NO: 2 over its entire length, and still more preferably atleast 90% identity, and even still more preferably at least 95% identityto SEQ ID NO: 2. Furthermore, those with at least 97-99% are highlypreferred. Preferably TR7 polypeptides exhibit at least one biologicalactivity of the receptor.

The TR7 polypeptides may be in the form of the “mature” protein or maybe a part of a larger protein such as a fusion protein. It is oftenadvantageous to include an additional amino acid sequence which containssecretory or leader sequences, pro-sequences, sequences which aid inpurification such as multiple histidine residues, or an additionalsequence for stability during recombinant production.

Fragments of the TR7 polypeptides are also included in the invention. Afragment is a polypeptide having an amino acid sequence that entirely isthe same as part, but not all, of the amino acid sequence of theaforementioned TR7 polypeptides. As with TR7 polypeptides, fragments maybe “free-standing,” or comprised within a larger polypeptide of whichthey form a part or region, most preferably as a single continuousregion. Representative examples of polypeptide fragments of theinvention, include, for example, fragments from about amino acid number1-20, 21-40, 41-60, 61-80, 81-100, and 101 to the end of TR7polypeptide. In this context “about” includes the particularly recitedranges larger or smaller by several, 5, 4, 3, 2 or 1 amino acid ateither extreme or at both extremes.

Preferred fragments include, for example, truncation polypeptides havingthe amino acid sequence of TR7 polypeptides, except for deletion of acontinuous series of residues that includes the amino terminus, or acontinuous series of residues that includes the carboxyl terminus ordeletion of two continuous series of residues, one including the aminoterminus and one including the carboxyl terminus. Also preferred arefragments characterized by structural or functional attributes such asfragments that comprise alpha-helix and alpha-helix forming regions,beta-sheet and beta-sheet-forming regions, turn and turn-formingregions, coil and coil-forming regions, hydrophilic regions, hydrophobicregions, alpha amphipathic regions, beta amphipathic regions, flexibleregions, surface-forming regions, substrate binding region, and highantigenic index regions. Other preferred fragments are biologicallyactive fragments. Biologically active fragments are those that mediatereceptor activity, including those with a similar activity or animproved activity, or with a decreased undesirable activity. Alsoincluded are those that are antigenic or immunogenic in an animal,especially in a human.

Preferably, all of these polypeptide fragments retain the biologicalactivity of the receptor, including antigenic activity. Among the mostpreferred fragment is that having the amino acid sequence of SEQ ID NO:4. Variants of the defined sequence and fragments also form part of thepresent invention. Preferred variants are those that vary from thereferents by conservative amino acid substitutions—i.e., those thatsubstitute a residue with another of like characteristics. Typical suchsubstitutions are among Ala, Val, Leu and Ile; among Ser and Thr; amongthe acidic residues Asp and Glu; among Asn and Gln; and among the basicresidues Lys and Arg; or aromatic residues Phe and Tyr. Particularlyprefeed are variants in which several 5-10, 1-5, or 1-2 amino acids aresubstituted, deleted, or added in any combination.

The TR7 polypeptides of the invention can be prepared in any suitablemanner. Such polypeptides include isolated naturally occurringpolypeptides, recombinantly produced polypeptides, syntheticallyproduced polypeptides, or polypeptides produced by a combination ofthese methods. Means for preparing such polypeptides are well understoodin the art.

Polynucleotides of the Invention

Another aspect of the invention relates to TR7 polynucleotides. TR7polynucleotides include isolated polynucleotides which encode the TR7polypeptides and fragments, and polynucleotides closely related thereto.More specifically, TR7 polynucleotide of the invention include apolynucleotide comprising the nucleotide sequence contained in SEQ IDNO:1 encoding a TR7 polypeptide of SEQ ID NO:2, and polynucleotideshaving the particular sequences of SEQ ID NOS: 1 and 3. TR7polynucleotides further include a polynucleotide comprising a nucleotidesequence that has at least 80% identity over its entire length to anucleotide sequence encoding the TR7 polypeptide of SEQ ID NO:2, and apolynucleotide comprising a nucleotide sequence that is at least 80%identical to that of SEQ ID NO:1 over its entire length. In this rsuchasard, polynucleotides at least 90% identical are particularlypreferred, and those with at least 95% are especially preferred.Furthermore, those with at least 97% are highly preferred and those withat least 98-99% are most highly preferred, with at least 99% being themost preferred. Also included under TR7 polynucleotides are a nucleotidesequence which has sufficient identity to a nucleotide sequencecontained in SEQ ID NO:1 to hybridize under conditions useable foramplification or for use as a probe or marker. The invention alsoprovides polynucleotides which are complementary to such TR7polynucleotides.

TR7 of the invention is structurally related to other proteins of theTumor necrosis factor receptor (TNF-R) family, as shown by the resultsof sequencing the cDNA encoding human TR7. The cDNA sequence of SEQ IDNO:1 contains an open reading frame (nucleotide number 7 to 1974)encoding a polypeptide of 655 amino acids of SEQ ID NO:2. The amino acidsequence of Table 1 (SEQ ID NO:2) has about 35% identity (using BLAST(from GCG)) in 168 amino acid residues with human osteprotegerin (OPG)protein (Simonet W S, et al., Cell 89: 309-319 (1997)). The nucleotidesequence of Table 1 (SEQ ID NO:1) has about 65% identity (using BESTFIT(from GCG)) in 102 nucleotide residues with Human Herpvesvirus EntryMediator (HVEM) (Montgomery, R. I. et al., cell 87, 427 (1996)) and 57%identity (using BESTFIT (from GCG)) in 118 nucleotide residues withhuman Osteoprotegerin (OPG) Protein (Simonet W S, et al., Cell 89:309-319 (1997)). Thus TR7 polypeptides and polynucleotides of thepresent invention are expected to have, inter alia, similar biologicalfunctions/properties to their homologous polypeptides andpolynucleotides, and their utility is obvious to anyone skilled in theart.

TABLE 1^(a)    1 TCAGCCATGG GGACCTCTCC GAGCAGCAGC ACCGCCCTCG CCTCCTGCAG  51 CCGCATCGCC CGCCGAGCCA CAGCCACGAT GATCGCGGGC TCCCTTCTCC  101TGCTTGGATT CCTTAGCACC ACCACAGCTC AGCCAGAACA GAAGGCCTCG  151 AATCTCATTGGCACATACCG CCATGTTGAC CGTGCCACCG GCCAGGTGCT  201 AACCTGTGAC AAGTGTCCAGCAGGAACCTA TGTCTCTGAG CATTGTACCA  251 ACACAAGCCT GCGCGTCTGC AGCAGTTGCCCTGTGGGGAC CTTTACCAGG  301 CATGAGAATG GCATAGAGAA ATGCCATGAC TGTAGTCAGCCATGCCCATG  351 GCCAATGATT GAGAAATTAC CTTGTGCTGC CTTGACTGAC CGAGAATGCA 401 CTTGCCCACC TGGCATGTTC CAGTCTAACG CTACCTGTGC CCCCCATACG  451GTGTGTCCTG TGGGTTGGGG TGTGCGGAAG AAAGGGACAG AGACTGAGGA  501 TGTGCGGTGTAAGCAGTGTG CTCGGGGTAC CTTCTCAGAT GTGCCTTCTA  551 GTGTGATGAA ATGCAAAGCATACACAGACT GTCTGAGTCA GAACCTGGTG  601 GTGATCAAGC CGGGGACCAA GGAGACAGACAACGTCTGTG GCACACTCCC  651 GTCCTTCTCC AGCTCCACCT CACCTTCCCC TGGCACAGCCATCTTTCCAC  701 GCCCTGAGCA CATGGAAACC CATGAAGTCC CTTCCTCCAC TTATGTTCCC 751 AAAGGCATGA ACTCAACAGA ATCCAACTCT TCTGCCTCTG TTAGACCAAA  801GGTACTGAGT AGCATCCAGG AAGGGACAGT CCCTGACAAC ACAAGCTCAG  851 CAAGGGGGAAGGAAGACGTG AACAAGACCC TCCCAAACCT TCAGGTAGTC  901 AACCACCAGC AAGGCCCCCACCACAGACAC ATCCTGAAGC TGCTGCCGTC  951 CATGGAGGCC ACTGGGGGCG AGAAGTCCAGCACGCCCATC AAGGGCCCCA 1001 AGAGGGGACA TCCTAGACAG AACCTACACA AGCATTTTGACATCAATGAG 1051 CATTTGCCCT GGATGATTGT GCTTTTCCTG CTGCTGGTGC TTGTGGTGAT1101 TGTGGTGTGC AGTATCCGGA AAAGCTCGAG GACTCTGAAA AAGGGGCCCC 1151GGCAGGATCC CAGTGCCATT GTGGAAAAGG CAGGGCTGAA GAAATCCATG 1201 ACTCCAACCCAGAACCGGGA GAAATGGATC TACTACTGCA ATGGCCATGG 1251 TATCGATATC CTGAAGCTTGTAGCAGCCCA AGTGGGAAGC CAGTGGAAAG 1301 ATATCTATCA GTTTCTTTGC AATGCCAGTGAGAGGGAGGT TGCTGCTTTC 1351 TCCAATGGGT ACACAGCCGA CCACGAGCGG GCCTACGCAGCTCTGCAGCA 1401 CTGGACCATC CGGGGCCCCG AGGCCAGCCT CGCCCAGCTA ATTAGCGCCC1451 TGCGCCAGCA CCGGAGAAAC GATGTTGTGG AGAAGATTCG TGGGCTGATG 1501GAAGACACCA CCCAGCTGGA AACTGACAAA CTAGCTCTCC CGATGAGCCC 1551 CAGCCCGCTTAGCCCGAGCC CCATCCCCAG CCCCAACGCG AAACTTGAGA 1601 ATTCCGCTCT CCTGACGGTGGAGCCTTCCC CACAGGACAA GAACAAGGGC 1651 TTCTTCGTGG ATGAGTCGGA GCCCCTTCTCCGCTGTGACT CTACATCCAG 1701 CGGCTCCTCC GCGCTGAGCA GGAACGGTTC CTTTATTACCAAAGAAAAGA 1751 AGGACACAGT GTTGCGGCAG GTACGCCTGG ACCCCTGTGA CTTGCAGCCT1801 ATCTTTGATG ACATGCTCCA CTTTCTAAAT CCTGAGGAGC TGCGGGTGAT 1851TGAAGAGATT CCCCAGGCTG AGGACAAACT AGACCGGCTA TTCGAAATTA 1901 TTGGAGTCAAGAGCCAGGAA GCCAGCCAGA CCCTCCTGGA CTCTGTTTAT 1951 AGCCATCTTC CTGACCTGCTGTAGAACATA GGGATACTGC ATTCTGGAAA 2001 TTACTCAATT TAGTGGCAGG GTGGTTTTTTAATTTTCTTC TGTTTCTGAT 2051 TTTTGTTGTT TGGGGTGTGT GTGTGTGTTT GTGTGTGTGTGTGTGTGTGT 2101 GTGTGTGTGT GTTTAACAGA GAAAATGGGC AGTGCTTGAA TTCTTTCTCC2151 TTCTCTCTCT CTCTTTTTTT TTTAAATAAC TCCTCT ^(a)A nucleotide sequenceof a human TR7 (SEQ ID NO: 1).

TABLE 2^(b)   1 MGTSPSSSTA LASCSRIARR ATATMIAGSL LLLGFLSTTT AQPEQKASNL 51 IGTYRHVDRA TGQVLTCDKC PAGTYVSEHC TNTSLRVCSS CPVGTFTRHE 101NGIEKCHDCS QPCPWPMIEK LPCAALTDRE CTCPPGMFQS NATCAPHTVC 151 PVGWGVRKKGTETEDVRCKQ CARGTFSDVP SSVMKCKAYT DCLSQNLVVI 201 KPGTKETDNV CGTLPSFSSSTSPSPGTAIF PRPEHMETHE VPSSTYVPKG 251 MNSTESNSSA SVRPKVLSSI QEGTVPDNTSSARGKEDVNK TLPNLQVVNH 301 QQGPHHRHIL KLLPSMEATG GEKSSTPIKG PKRGHPRQNLHKHFDINEHL 351 PWMIVLFLLL VLVVIVVCSI RKSSRTLKKG PRQDPSAIVE KAGLKKSMTP401 TQNREKWIYY CNGHGIDILK LVAAQVGSQW KDIYQFLCNA SEREVAAFSN 451GYTADHERAY AALQHWTIRG PEASLAQLIS ALRQHRRNDV VEKIRGLMED 501 TTQLETDKLALPMSPSPLSP SPIPSPNAKL ENSALLTVEP SPQDKNKGFF 551 VDESEPLLRC DSTSSGSSALSRNGSFITKE KKDTVLRQVR LDPCDLQPIF 601 DDMLHFLNPE ELRVIEEIPQ AEDKLDRLFEIIGVKSQEAS QTLLDSVYSH 651 LPDLL* ^(b)An amino acid sequence of a humanTR7 (SEQ ID NO: 2)

One polynucleotide of the present invention encoding TR7 may be obtainedusing standard cloning and screening, from a cDNA library derived frommRNA in cells of human brain, heart, lung, thymus, kidney, smallintestine, prostate, monocytes and endothelial cells, using theexpressed sequence tag (EST) analysis (Adams, M. D., et al. Science(1991) 252:1651-1656; Adams, M. D. et al., Nature, (1992) 355:632-634;Adams, M. D., et al., Nature (1995) 377 Supp:3-174). Polynucleotides ofthe invention can also be obtained from natural sources such as genomicDNA libraries or can be synthesized using well known and commerciallyavailable techniques.

The nucleotide sequence encoding TR7 polypeptide of SEQ ID NO:2 may beidentical to the polypeptide encoding sequence contained in Table 1(nucleotide number 7 to 1974 of SEQ ID NO:1), or it may be a sequence,which as a result of the redundancy (degeneracy) of the genetic code,also encodes the polypeptide of SEQ ID NO:2.

When the polynucleotides of the invention are used for the recombinantproduction of TR7 polypeptide, the polynucleotide may include the codingsequence for the mature polypeptide or a fragment thereof, by itself,the coding sequence for the mature polypeptide or fragment in readingframe with other coding sequences, such as those encoding a leader orsecretory sequence, a pre-, or pro- or prepro-protein sequence, or otherfusion peptide portions. For example, a marker sequence whichfacilitates purification of the fused polypeptide can be encoded. Incertain preferred embodiments of this aspect of the invention, themarker sequence is a hexa-histidine peptide, as provided in the pQEvector (Qiagen, Inc.) and described in Gentz et al., Proc Natl Acad SciUSA (1989) 86:821-824, or is an HA tag. The polynucleotide may alsocontain non-coding 5′ and 3′ sequences, such as transcribed,non-translated sequences, splicing and polyadenylation signals, ribosomebinding sites and sequences that stabilze mRNA.

Further preferred embodiments are polynucleotides encoding TR7 variantscomprising the amino acid sequence of TR7 polypeptide of Table 2 (SEQ IDNO:2) in which several, 5-10, 1-5, 1-3, 1-2 or 1 amino acid residues aresubstituted, deleted or added, in any combination. Among the preferredpolynucleotides of the present invention is contained in Table 3 (SEQ IDNO: 3) encoding the amino acid sequence of Table 4 (SEQ ID NO: 4).

TABLE 3^(c)    1 GCGNCCGCGN NGNGNGCAAG GTGCTGAGCG CCCCTAGNGC CTCCCTTGCC  51 GCCTCCCTCC TCTGCCCGGC CGTAGCAGTG CACATGGGGT GTTGGAGGTA  101GATGGGCTCC CGGCCGGGAG GCGGCGGTGG ATGCGGCGCT GGGCAGAAGC  151 AGCCGCCGATTCCAGCTGCC CCGCGCGCCC CGGCCACCTT GCGAGTCCCC  201 GGTTCAGCCA TGGGGACCTCTCCGAGCAGC AGCACCGCCC TCGCCTCCTG  251 CAGCCGCATC GCCCGCCGAG CCACAGCCACGATGATCGCG GGCTCCCTTC  301 TCCTGCTTGG ATTCCTTAGC ACCACCACAG CTCAGCCAGAACAGAAGGCC  351 TCGAATCTCA TTGGCACATA CCGCCATGTT GACCGTGCCA CCGGCCAGGT 401 GCTAACCTGT GACAAGTGTC CAGCAGGAAC CTATGTCTCT GAGCATTGTA  451CCAACACAAG CCTGCGCGTC TGCAGCAGTT GCCCTGTGGG GACCTTTACC  501 AGGCATGAGAATGGCATAGA GAAATGCCAT GACTGTAGTC AGCCATGCCC  551 ATGGCCAATG ATTGAGAAATTACCTTGTGC TGCC ^(c)A partial nucleotide sequence of a humanTR7 (SEQ IDNO: 3).

TABLE 4^(d)   1 MGTSPSSSTA LASCSRIARR ATATMIAGSL LLLGFLSTTT AQPEQKASNL 51 IGTYRHVDRA TGQVLTCDKC PAGTYVSEHC TNTSLRVCSS CPVGTFTRHE 101NGIEKCHDCS QPCPWPMIEK LPCAA ^(d)A partial amino acid sequence of a humanTR7 (SEQ ID NO: 4).

The present invention further relates to polynucleotides that hybridizeto the herein above-describes sequences. In this regard, the presentinvention especially relates to polynucleotides which hybridize understringent conditions to the herein above-described polynucleotides. Asherein used, the term “stringent conditions” means hybridization willoccur only if there is at least 80%, and preferably at least 90%, andmore preferably at least 95%, yet even more preferably 97-99% identitybetween the sequences.

Polynucleotides of the invention, which are identical or sufficientlyidentical to a nucleotide sequence contained in SEQ ID NO:1 or afragment thereof, may be used as hybridization probes for cDNA andgenomic DNA, to isolate fill-length cDNAs and genomic clones encodingTR7 and to isolate cDNA and genomic clones of other genes (includinggenes encoding homologs and orthologs from species other than human)that have a high sequence similarity to the TR7 gene. Such hybridizationtechniques are known to those of skill in the art. Typically thesenucleotide sequences are 80% identical, preferably 90% identical, morepreferably 95% identical to that of the referent. The probes generallywill comprise at least 15 nucleotides. Preferably, such probes will haveat least 30 nucleotides and may have at least 50 nucleotides.Particularly preferred probes will range between 30 and 50 nucleotides.

In one embodiment, to obtain a polynucleotide encoding TR7 polypeptide,including homologs and orthologs from species other than human,comprises the steps of screening an appropriate library under stingenthybridization conditions with a labeled probe having the SEQ ID NO: 1 ora fragment thereof (including that of SEQ ID NO: 3), and isolatingfull-length cDNA and genomic clones containing said polynucleotidesequence. Such hybridization techniques are well known to those of skillin the art. Stringent hybridization conditions are as defined above oralternatively conditions under overnight incubation at 42° C. in asolution comprising 50% formamide, 5×SSC (150 mM NaCl, 15 mM trisodiumcitrate), 50 mM sodium phosphate (pH7.6), 5×Denhardt's solution, 10%dextran sulfate, and 20 microgram/ml denatured, sheared salmon spermDNA, followed by washing the filters in 0.1×SSC at about65° C.

The polynucleotides and polypeptides of the present invention may beemployed as research reagents and materials for discovery of treatmentsand diagnostics to animal and human disease.

Vectors, Host Cells, Expression

The present invention also relates to vectors which comprise apolynucleotide or polynucleotides of the present invention, and hostcells which are genetically engineered with vectors of the invention andto the production of polypeptides of the invention by recombinanttechniques. Cell-free translation systems can also be employed toproduce such proteins using RNAs derived from the DNA constructs of thepresent invention.

For recombinant production, host cells can be genetically engineered toincorporate expression systems or portions thereof for polynucleotidesof the present invention. Introduction of polynucleotides into hostcells can be effected by methods described in many standard laboratorymanuals, such as Davis et al., BASIC METHODS IN MOLECULAR BIOLOGY (1986)and Sambrook et al., MOLECULAR CLONING: A LABORATORY MANUAL, 2nd Ed.,Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y. (1989)such as calcium phosphate transfection, DEAE-dextran mediatedtransfection, transvection, microinjection, cationic lipid-mediatedtransfection, electroportion, transduction, scrape loading, ballisticintroduction or infection.

Representative examples of appropriate hosts include bacterial cells,such as streptococci, staphylococci, E. coli, Streptomyces and Bacillussubtilis cells; fungal cells, such as yeast cells and Aspergillus cells;insect cells such as Drosophila S2 and Spodoptera Sf9 cells; animalcells such as CHO, COS, HeLa, C127, 3T3, BHK, HEK 293 and Bowes melanomacells; and plant cells.

A great variety of expression systems can be used. Such systems include,among others, chromosomal, episomal and virus-derived systems, e.g.,vectors derived from bacterial plasmids, from bacteriophage, fromtransposons, from yeast episomes, from insertion elements, from yeastchromosomal elements, from viruses such as baculoviruses, papovaviruses, such as SV40, vaccinia viruses, adenoviruses, fowl pox viruses,pseudorabies viruses and retrovirses, and vectors derived fromcombinations thereof, such as those derived from plasmid andbacteriophage genetic elements, such as cosmids and phagemids. Theexpression systems may contain control regions that regulate as well asengender expression. Generally, any system or vector suitable tomaintain, propagate or express polynucleotides to produce a polypeptidein a host may be used. The appropriate nucleotide sequence may beinserted into an expression system by any of a variety of well-known androutine techniques, such as, for example, those set forth in Sambrook etal., MOLECULAR CLONING, A LABORATORY MANUAL (supra).

For secretion of the translated protein into the lumen of theendoplasmic reticulum, into the periplasmic space or into theextracellular environment, appropriate secretion signals may beincorporated into the desired polypeptide. These signals may beendogenous to the polypeptide or they may be heterologous signals.

If the TR7 polypeptide is to be expressed for use in screening assays,generally, it is preferred that the polypeptide be produced at thesurface of the cell. In this event, the cells may be harvested prior touse in the screening assay. If TR7 polypeptide is secreted into themedium, the medium can be recovered in order to recover and purify thepolypeptide; if produced intracellularly, the cells must first be lysedbefore the polypeptide is recovered.

TR7 polypeptides can be recovered and purified from recombinant cellcultures by well-known methods including ammonium sulfate or ethanolprecipitation, acid extraction, anion or cation exchange chromatography,phosphocellulose chromatography, hydrophobic interaction chromatography,affinity chromatography, hydroxylapatite chromatography and lectinchromatography. Most preferably, high performance liquid chromatographyis employed for purification. Well known techniques for refoldingproteins may be employed to regenerate active conformation when thepolypeptide is denatured during isolation and or purification.

Diagnostic Assays

This invention also relates to the use of TR7 polynucleotides for use asdiagnostic reagents. Detection of a mutated form of TR7 gene associatedwith a dysfunction will provide a diagnostic tool that can add to ordefine a diagnosis of a disease or susceptibility to a disease whichresults from under-expression, over-expression or altered expression ofTR7. Individuals carrying mutations in the TR7 gene may be detected atthe DNA level by a variety of techniques.

Nucleic acids for diagnosis may be obtained from a subject's cells, suchas from blood, urine, saliva, tissue biopsy or autopsy material. Thegenomic DNA may be used directly for detection or may be amplifiedenzymatically by using PCR or other amplification techniques prior toanalysis. RNA or cDNA may also be used in similar fashion. Deletions andinsertions can be detected by a change in size of the amplified productin comparison to the normal genotype. Point mutations can be identifiedby hybridizing amplified DNA to labeled TR7 nucleotide sequences.Perfectly matched sequences can be distinguished from mismatchedduplexes by RNase digestion or by differences in melting temperaturesDNA sequence differences may also be detected by alterations inelectrophoretic mobility of DNA fragments in gels, with or withoutdenaturing agents, or by direct DNA sequencing. See, e.g., Myers et al.,Science (1985) 230:1242. Sequence changes at specific locations may alsobe revealed by nuclease protection assays, such as RNase and S1protection or the chemical cleavage method. See Cotton et al., Proc NatlAcad Sci USA (1985) 85: 4397-4401. In another embodiment, an array ofoligonucleotides probes comprising TR7 nucleotide sequence or fragmentsthereof can be constructed to conduct efficient screening of e.g.,genetic mutations. Array technology methods are well known and havegeneral applicability and can be used to address a variety of questionsin molecular genetics including gene expression, genetic linkage, andgenetic variability. (See for example: M. Chee et al., Science, Vol 274,pp 610-613 (1996)).

The diagnostic assays offer a process for diagnosing or determining asusceptibility to chronic and acute inflammation, arthritis, septicemiaautoimmune diseases (such as inflammatory bowel disease and psoriasis),transplant rejection, graft versus host disease, infection, stroke,ischemia, acute respiratory disease syndrome, restenosis, brain injury,AIDS, bone diseases, cancer (such as lymphoproliferative disorders),atherosclerosis, and Alzheimers disease through detection of mutation inthe TR7 gene by the methods described.

In addition, chronic and acute inflammation, arthritis, septicemia,autoimmune diseases (such as inflammatory bowel disease and psoriasis),transplant rejection, graft versus host disease, infection, stroke,ischemia, acute respiratory disease syndrome, restenosis, brain injury,AIDS, bone diseases, cancer (such as lymphoproliferative disorders),atherosclerosis, and Alzheimers disease, can be diagnosed by methodscomprising determining from a sample derived from a subject anabnormally decreased or increased level of TR7 polypeptide or TR7 mRNA.Decreased or increased expression can be measured at the RNA level usingany of the methods well known in the art for the quantitation ofpolynucleotides, such as, for example, PCR, RT-PCR, RNase protection,Northern blotting and other hybridization methods. Assay techniques thatcan be used to determine levels of a protein, such as an TR7, in asample derived from a host are well-known to those of skill in the art.Such assay methods include radioimmunoassays, competitive-bindingassays, Western Blot analysis and ELISA assays.

Thus in another aspect, the present invention relates to a diagonostickit for a disease or suspectability to a disease, particularly chronicand acute inflammation, arthritis, septicemia, autoimmune diseases (suchas inflammatory bowel disease and psoriasis), transplant rejection,graft versus host disease, infection, stroke, ischemia, acuterespiratory disease syndrome, restenosis, brain injury, AIDS, bonediseases, cancer (such as lymphoproliferative disorders),atherosclerosis, and Alzheimers disease, which comprises:

(a) a TR7 polynucleotide, preferably the nucleotide sequence of SEQ IDNO: 1, or a fragment thereof;

(b) a nucleotide sequence complementary to that of (a);

(c) a TR7 polypeptide, preferably the polypeptide of SEQ ID NO: 2, or afragment thereof; or

(d) an antibody to a TR7 polypeptide, preferably to the polypeptide ofSEQ ID NO: 2.

It will be appreciated that in any such kit, (a), (b), (c) or (d) maycomprise a substantial component.

Chromosome Assays

The nucleotide sequences of the present invention are also valuable forchromosome identification. The sequence is specifically targeted to andcan hybridize with a particular location on an individual humanchromosome. The mapping of relevant sequences to chromosomes accordingto the present invention is an important first step in correlating thosesequences with gene associated disease. Once a sequence has been mappedto a precise chromosomal location, the physical position of the sequenceon the chromosome can be correlated with genetic map data. Such data arefound, for example, in V. McKusick, Mendelian Inheritance in Man(available on fine through Johns Hopkins University Welch MedicalLibrary). The relationship between genes and diseases that have beenmapped to the same chromosomal region are then identified through imageanalysis (coinheritance of physically adjacent genes). The differencesin the cDNA or genomic sequence between affected and unaffectedindividuals can also be determined. If a mutation is observed in some orall of the affected individuals but not in any normal individuals, thenthe mutation is likely to be the causative agent of the disease.

Antibodies

The polypeptides of the invention or ther fragments or analogs thereofor cells expressing them can also be used as immunogens to produceantibodies immunospecific for the TR7 polypeptides. The term“immunospecific” means that the antibodies have substantiall greateraffinity for the polypeptides of the invention than their affinity forother related polypeptides in the prior art.

Antibodies generated against the TR7 polypeptides can be obtained byadministering the polypeptides or epitope-bearing fragments, analogs orcells to an animal, preferably a nonhuman, using routine protocols. Forpreparation of monoclonal antibodies, any technique which providesantibodies produced by continuous cell line cultures can be used.Examples include the hybridoma technique Kohler, G. and Milstein, C.,Nature (1975) 256:495-497), the trioma technique, the human B-cellhybridoma technique (Kozbor et al., Immunology Today (1983) 4:72) andthe EBV-hybridoma technique (Cole et al., MONOCLONAL ANTIBODIES ANDCANCER THERAPY, pp. 77-96, Alan R Liss, Inc., 1985).

Techniques for the production of single chain antibodies (U.S. Pat. No.4,946,778) can also be adapted to produce single chain antibodies topolypeptides of this invention. Also, transgenic mice, or otherorganisms including other mammals, may be used to express humanizedantibodies.

The above-described antibodies may be employed to isolate or to identifyclones expressing the polypeptide or to purify the polypeptides byaffinity chromatography.

Antibodies against TR7 polypeptides may also be employed to treatchronic and acute inflammation, arthritis, septicemia, autoimmunediseases (such as inflammatory bowel disease and psoriasis), transplantrejection, graft versus host disease, infection, stroke, ischemia, acuterespiratory disease syndrome, restenosis, brain injury, AIDS, bonediseases, cancer (such as lymphoproliferative disorders),atherosclerosis, and Alzheimers disease, among others.

Vaccines

Another aspect of the invention relates to a method for inducing animmunological response in a mammal which comprises inoculating themammal with TR7 polypeptide, or a fragment thereof, adequate to produceantibody and/or T cell immune response to protect said animal fromchronic and acute inflammation, arthritis, septicemia, autoimmunediseases (such as inflammatory bowel disease and psoriasis), transplantrejection, graft versus host disease, infection, stroke, ischemia, acuterespiratory disease syndrome, restenosis, brain injury, AIDS, bonediseases, cancer (such as lymphoproliferative disorders),atherosclerosis, and Alzheimers disease, among others. Yet anotheraspect of the invention relates to a method of inducing immunologicalresponse in a mammal which comprises, delivering TR7 polypeptide via avector directing expression of TR7 polynucleotide in vivo in order toinduce such an immunological response to produce antibody to protectsaid animal from diseases.

Further aspect of the invention relates to an immunological/vaccineformulation (composition) which, when introduced into a mammalian host,induces an immunological response in that mammal to a TR7 polypeptidewherein the composition comprises a TR7 polypeptide or TR7 gene. Thevaccine formulation may further comprise a suitable carrier. Since TR7polypeptide may be broken down in the stomach, it is preferablyadministered parenterally (including subcutaneous, intramuscular,intravenous, intradermal etc. injection). Formulations suitable forparenteral administration include aqueous and non-aqueous sterileinjection solutions which may contain anti-oxidants, buffers,bacteriostats and solutes which render the formulation instonic with theblood of the recipient; and aqueous and non-aqueous sterile suspensionswhich may include suspending agents or thickening agents. Theformulations may be presented in unit-dose or multi-dose containers, forexample, sealed ampoules and vials and may be stored in a freeze-driedcondition requiring only the addition of the sterile liquid carrierimmediately prior to use. The vaccine formulation may also includeadjuvant systems for enhancing the immunogenicity of the formulation,such as oil-in water systems and other systems known in the art. Thedosage will depend on the specific activity of the vaccine and can bereadily determined by routine experimentation.

Screening Assays

The TR7 polypeptide of the present invention may be employed in ascreening process for compounds which bind the receptor and whichactivate (agonists) or inhibit activation of (antagonists) the receptorpolypeptide of the present invention. Thus, polypeptides of theinvention may also be used to assess the binding of small moleculesubstrates and ligands in, for example, cells, cell-free preparations,chemical libraries, and natural product mixtures. These substrates andligands may be natural substrates and ligands or may be structural orfunctional mimetics. See Coligan et al., Current Protocols in Immunology1(2):Chapter 5 (1991).

TR7 polypeptides are responsible for many biological functions,including many pathologies. Accordingly, it is desirous to findcompounds and drugs which stimulate TR7 on the one hand and which caninhibit the function of TR7 on the other hand. In general, agonists areemployed for therapeutic and prophylactic purposes for such conditionsas chronic and acute inflammation, arthritis, septicemia, autoimmunediseases (such as inflammatory bowel disease and psoriasis), transplantrejection, graft versus host disease, infection, stroke, ischemia, acuterespiratory disease syndrome, restenosis, brain injury, AIDS, bonediseases, cancer (such as lymphoproliferative disorders),atherosclerosis, and Alzheimers disease. Antagonists may be employed fora variety of therapeutic and prophylactic purposes for such conditionsas chronic and acute inflammation, arthritis, septicemia, autoimmunediseases (such as inflammatory bowel disease and psoriasis), transplantrejection, graft versus host disease, infection, stroke, ischemia, acuterespiratory disease syndrome, restenosis, brain injury, AIDS, bonediseases, cancer (such as lymphoproliferative disorders),atherosclerosis, and Alzheimers disease.

In general, such screening procedures involve producing appropriatecells which express the receptor polypeptide of the present invention onthe surface thereof. Such cells include cells from mammals, yeast,Drosophila or E. coli. Cells expressing the receptor (or cell membranecontaining the expressed receptor) are then contacted with a testcompound to observe binding, or stimulation or inhibition of afunctional response.

The assays may simply test binding of a candidate compound whereinadherence to the cells bearing the receptor is detected by means of alabel directly or indirectly associated with the candidate compound orin an assay involving competition with a labeled competitor. Further,these assays may test whether the candidate compound results in a signalgenerated by activation of the receptor, using detection systemsappropriate to the cells bearing the receptor at their surfaces.Inhibitors of activation are generally assayed in the presence of aknown agonist and the effect on activation by the agonist by thepresence of the candidate compound is observed.

Further, the assays may simply comprise the steps of mixing a candidatecompound with a solution containing a TR7 polypeptide to form a mixture,measuring TR7 activity in the mixture, and comparing the TR7 activity ofthe mixture to a standard.

The TR7 cDNA, protein and antibodies to the protein may also be used toconfigure assays for detecting the effect of added compounds on theproduction of TR7 mRNA and protein in cells. For example, an ELISA maybe constructed for measuring secreted or cell associated levels of TR7protein using monoclonal and polyclonal antibodies by standard methodsknown in the art, and this can be used to discover agents which mayinhibit or enhance the production of TR7 (also called antagonist oragonist, respectively) from suitably manipulated cells or tissues.Standard methods for conducting screening assays are well understood inthe art.

Examples of potential TR7 antagonists include antibodies or, in somecases, oligonucleotides or proteins which are closely related to theligand of the TR7, e.g., a fragment of the ligand, or small moleculeswhich bind to the receptor but do not elicit a response, so that theactivity of the receptor is prevented.

Thus in another aspect, the present invention relates to a screening kitfor identifying agonists, antagonists, ligands, receptors, substrates,enzymes, etc. for TR7 polypeptides; or compounds which decrease orenhance the production of TR7 polypeptides, which comprises:

(a) a TR7 polypeptide, preferably that of SEQ ID NO:2;

(b) a recombinant cell expressing a TR7 polypeptide, preferably that ofSEQ ID NO:2;

(c) a cell membrane expressing a TR7 polypeptide; preferably that of SEQID NO: 2; or

(d) antibody to a TR7 polypeptide, preferably that of SEQ ID NO: 2.

It will be appreciated that in any such kit, (a), (b), (c) or (d) maycomprise a substantial component.

Prophylactic and Therapeutic Methods

This invention provides methods of treating an abnormal conditionsrelated to both an excess of and insufficient amounts of TR7 activity.

If the activity of TR7 is in excess, several approaches are available.One approach comprises administering to a subject an inhibitor compound(antagonist) as hereinabove described along with a pharmaceuticallyacceptable carrier in an amount effective to inhibit activation byblocking binding of ligands to the TR7, or by inhibiting a secondsignal, and thereby alleviating the abnormal condition.

In another approach, soluble forms of TR7 polypeptides still capable ofbinding the ligand in competition with endogenous TR7 may beadministered. Typical embodiments of such competitors comprise fragmentsof the TR7 polypeptide.

In still another approach, expression of the gene encoding endogenousTR7 can be inhibited using expression blocking techniques. Known suchtechniques involve the use of antisense sequences, either internallygenerated or separately administered. See, for example, O'Connor, JNeurochem (1991) 56:560 in Oligodeoxynucleotides as Antisense Inhibitorsof Gene Expression, CRC Press, Boca Raton, Fla. (1988). Alternatively,oligonucleotides which form triple helices with the gene can besupplied. See, for example, Lee et al., Nucleic Acids Res (1979) 6:3073;Cooney et al., Science (1988) 241:456; Dervan et al., Science (1991)251:1360. These oligomers can be administered per se or the relevantoligomers can be expressed in vivo.

For treating abnormal conditions related to an under-expression of TR7and its activity, several approaches are also available. One approachcomprises administering to a subject a therapeutically effective amountof a compound which activates TR7, i.e., an agonist as described above,in combination with a pharmaceutically acceptable carrier, to therebyalleviate the abnormal condition. Alternatively, gene therapy may beemployed to effect the endogenous production of TR7 by the relevantcells in the subject. For example, a polynucleotide of the invention maybe engineered for expression in a replication defective retroviralvector, as discussed above. The retroviral expression construct may thenbe isolated and introduced into a packaging cell transduced with aretroviral plasmid vector containing RNA encoding a polypeptide of thepresent invention such that the packaging cell now produces infectiousviral particles containing the gene of interest. These producer cellsmay be administered to a subject for engineering cells in viva andexpression of the polypeptide in vivo. For overview of gene therapy, seeChapter 20, Gene Therapy and other Molecular Genetic-based TherapeuticApproaches, (and references cited therein) in Human Molecular Genetics,T Strachan and A P Read, BIOS Scientific Publishers Ltd (1996).

Formulation and Administration

Peptides, such as the soluble form of TR7 polypeptides, and agonists andantagonist peptides or small molecules, may be formulated in combinationwith a suitable pharmaceutical carrier. Such formulations comprise atherapeutically effective amount of the polypeptide or compound, and apharmaceutically acceptable carrier or excipient. Such carriers includebut are not limits to, saline, buffered saline, dextrose, water,glycerol ethanol, and combinations thereof. Formulation should suit themode of administration, and is well within the skill of the art. Theinvention further relates to pharmaceutical packs and kits comprisingone or more containers filled with one or more of the ingredients of theaforementioned compositions of the invention.

Polypeptides and other compounds of the present invention may beemployed alone or in conjunction with other compounds, such astherapeutic compounds.

Preferred forms of systemic administration of the pharmaceuticalcompositions include injection, typically by intravenous injection.Other injection routes, such as subcutaneous, intramuscular, orintraperitoneal, can be used. Alternative means for systemicadministration include transmucosal and transdermal administration usingpenetrants such as bile salts or fusidic acids or other detergents. Inaddition, if properly formulated in enteric or encapsulatedformulations, oral administration may also be possible. Administrationof these compounds may also be topical and/or localized, in the form ofsalves, pastes, gels and the like.

The dosage range required is on the choice of peptide, the route ofadministration, the nature of the formulation, the nature of thesubject's condition, and the judgment of the attending practitioner.Suitable dosages, however, are in the range of 0.1-100 μg/kg of subject.Wide variations in the needed dosage, however, are to be expected inview of the variety of compounds available and the differingefficiencies of various routes of administration. For example, oraladministration would be expected to require higher dosages thanadministration by intravenous injection. Variations in these dosagelevels can be adjusted using standard empirical routines foroptimization, as is well understood in the art.

Polypeptides used in treatment can also be generated endogenously in thesubject, in treatment modalities often referred to as “gene therapy” asdescribed above. Thus, for example, cells from a subject may beengineered with a polynucleotide, such as a DNA or RNA, to encode apolypeptide ex vivo, and for example, by the use of a retroviral plasmidvector. The cells are then introduced into the subject.

EXAMPLE 1

Three ESTs (EST#1502886,EST#9843791 and EST#2051015) with sequencesimilarity to the human TNF receptor were discovered in a commercial ESTdatabase. Analysis of two (EST#843791 and EST#2051015) nucleotidesequences, revealed that each was a partial sequence of the completecoding sequence, overlapping, with 100% identity, 108 bp at thenucleotide level. Together, the two sequences encompassed the completepredicted coding sequence of 1,968 bp, and encoded an open reading framefor a novel member of the TNF receptor superfamily and named TR7. Thepredicted protein is 655 amino acids long with a hydrophobic membranespanning region indicating that at least one form of TR7 is expressed asa membrane-bound protein. Comparison of TR7 protein sequence, with otherTNF receptor family proteins indicates that it has four of thecysteine-rich repeats which are characteristic of the extracellulardomains of this family, and of an intracellular death domain.

All publications, including but not limited to patents and patentapplications, cited in this specification are herein incorporated byreference as if each individual publication were specifically andindividually indicated to be incorporated by reference herein as thoughfully set forth.

TABLE 3 Nucleotide and Amino Acid sequence of a TR7 fragment (SEQ IDNOS: 5 and 6, respectively.)   1GCGNCCGCGNNGNGNGCAAGGTGCTGAGCGCCCCTAGNGCCTCCCTTGCCGCCTCCCTCC  60  61TCTGCCCGGCCGTAGCAGTGCACATGGGGTGTTGGAGGTAGATGGGCTCCCGGCCGGGAG 120 121GCGGCGGTGGATGCGGCGCTGGGCAGAAGCAGCCGCCGATTCCAGCTGCCCCGCGCGCCC 180 181CGGCCACCTTGCGAGTCCCCGGTTCAGCCATGGGGACCTCTCCGAGCAGCAGCACCGCCC 240 241TCGGCCTCCTGCAACCGCATCGCCCGCCGAGCCACAGCCACGATGATCGCGGGCTCCCTT 300                                          MetIleAlaGlySerLeu   6 301CTCCTGCTTGGATTCCTTAGCACCACCACAGCTCAGCCAGAACAGAAGGCCTCGAATCTC 360   7LeuLeuLeuGlyPheLeuSerThrThrThrAlaGlnProGluGlnLysAlaSerAsnLeu  26 361ATTGGCACATACCGCCATGTTGACCGTGCCACCGGCCAGGTGCTAACCTGTGACAAGTGT 420  27IleGlyThrTyrArgHisValAspArgAlaThrGlyGlnValLeuThrCysAspLysCys  46 421CCAGCAGGAACCTATGTCTCTGAGCATTGTACCAACACAAGCCTGCGCGTCTGTCAGCAG 480  47ProAlaGlyThrTyrValSerGluHisCysThrAsnThrSerLeuArgValCysGlnGln  66 481TGCCCTGTGGGGACCTTTACCAGGCATGAGAATGGCATAGAGAAATGCCATGACTGTAGT 540  67CysProValGlyThrPheThrArgHisGluAsnGlyIleGluLysCysHisAspCysSer  86 541CAGCCATGCCCATGGCCAATGATTGAGAAATTACCTTGTGCCTCTGCC 588  87GlnProCysProTrpProMetIleGluLysLeuProCysAlaSerAla 102

EXAMPLE 2

An EST (EST#1502886; Project ID: HHFGD57) with sequence similarity tothe human TNF receptor was discovered in a commercial EST database.Analysis of the 588 nucleotide sequence of the partial cDNA, indicatedthat it encoded an open reading frame for a novel member of the TNFreceptor superfamily and was named TR7. The predicted partial proteinsequence is 102 amino acids long, with a hydrophobic amino-terminalleader sequence indicating that TR7 is expressed as a secreted or cellsurface membrane bound protein. Comparison of the TR7 partial proteinsequence with other TNF receptor family proteins indicates that it hasat least one of the cysteine-rich repeats characteristic of theextracellular domains of this family.

6 1 2186 DNA HOMO SAPIENS 1 tcagccatgg ggacctctcc gagcagcagc accgccctcgcctcctgcag ccgcatcgcc 60 cgccgagcca cagccacgat gatcgcgggc tcccttctcctgcttggatt ccttagcacc 120 accacagctc agccagaaca gaaggcctcg aatctcattggcacataccg ccatgttgac 180 cgtgccaccg gccaggtgct aacctgtgac aagtgtccagcaggaaccta tgtctctgag 240 cattgtacca acacaagcct gcgcgtctgc agcagttgccctgtggggac ctttaccagg 300 catgagaatg gcatagagaa atgccatgac tgtagtcagccatgcccatg gccaatgatt 360 gagaaattac cttgtgctgc cttgactgac cgagaatgcacttgcccacc tggcatgttc 420 cagtctaacg ctacctgtgc cccccatacg gtgtgtcctgtgggttgggg tgtgcggaag 480 aaagggacag agactgagga tgtgcggtgt aagcagtgtgctcggggtac cttctcagat 540 gtgccttcta gtgtgatgaa atgcaaagca tacacagactgtctgagtca gaacctggtg 600 gtgatcaagc cggggaccaa ggagacagac aacgtctgtggcacactccc gtccttctcc 660 agctccacct caccttcccc tggcacagcc atctttccacgccctgagca catggaaacc 720 catgaagtcc cttcctccac ttatgttccc aaaggcatgaactcaacaga atccaactct 780 tctgcctctg ttagaccaaa ggtactgagt agcatccaggaagggacagt ccctgacaac 840 acaagctcag caagggggaa ggaagacgtg aacaagaccctcccaaacct tcaggtagtc 900 aaccaccagc aaggccccca ccacagacac atcctgaagctgctgccgtc catggaggcc 960 actgggggcg agaagtccag cacgcccatc aagggccccaagaggggaca tcctagacag 1020 aacctacaca agcattttga catcaatgag catttgccctggatgattgt gcttttcctg 1080 ctgctggtgc ttgtggtgat tgtggtgtgc agtatccggaaaagctcgag gactctgaaa 1140 aaggggcccc ggcaggatcc cagtgccatt gtggaaaaggcagggctgaa gaaatccatg 1200 actccaaccc agaaccggga gaaatggatc tactactgcaatggccatgg tatcgatatc 1260 ctgaagcttg tagcagccca agtgggaagc cagtggaaagatatctatca gtttctttgc 1320 aatgccagtg agagggaggt tgctgctttc tccaatgggtacacagccga ccacgagcgg 1380 gcctacgcag ctctgcagca ctggaccatc cggggccccgaggccagcct cgcccagcta 1440 attagcgccc tgcgccagca ccggagaaac gatgttgtggagaagattcg tgggctgatg 1500 gaagacacca cccagctgga aactgacaaa ctagctctcccgatgagccc cagcccgctt 1560 agcccgagcc ccatccccag ccccaacgcg aaacttgagaattccgctct cctgacggtg 1620 gagccttccc cacaggacaa gaacaagggc ttcttcgtggatgagtcgga gccccttctc 1680 cgctgtgact ctacatccag cggctcctcc gcgctgagcaggaacggttc ctttattacc 1740 aaagaaaaga aggacacagt gttgcggcag gtacgcctggacccctgtga cttgcagcct 1800 atctttgatg acatgctcca ctttctaaat cctgaggagctgcgggtgat tgaagagatt 1860 ccccaggctg aggacaaact agaccggcta ttcgaaattattggagtcaa gagccaggaa 1920 gccagccaga ccctcctgga ctctgtttat agccatcttcctgacctgct gtagaacata 1980 gggatactgc attctggaaa ttactcaatt tagtggcagggtggtttttt aattttcttc 2040 tgtttctgat ttttgttgtt tggggtgtgt gtgtgtgtttgtgtgtgtgt gtgtgtgtgt 2100 gtgtgtgtgt gtttaacaga gaaaatgggc agtgcttgaattctttctcc ttctctctct 2160 ctcttttttt tttaaataac tcctct 2186 2 655 PRTHOMO SAPIENS 2 Met Gly Thr Ser Pro Ser Ser Ser Thr Ala Leu Ala Ser CysSer Arg 1 5 10 15 Ile Ala Arg Arg Ala Thr Ala Thr Met Ile Ala Gly SerLeu Leu Leu 20 25 30 Leu Gly Phe Leu Ser Thr Thr Thr Ala Gln Pro Glu GlnLys Ala Ser 35 40 45 Asn Leu Ile Gly Thr Tyr Arg His Val Asp Arg Ala ThrGly Gln Val 50 55 60 Leu Thr Cys Asp Lys Cys Pro Ala Gly Thr Tyr Val SerGlu His Cys 65 70 75 80 Thr Asn Thr Ser Leu Arg Val Cys Ser Ser Cys ProVal Gly Thr Phe 85 90 95 Thr Arg His Glu Asn Gly Ile Glu Lys Cys His AspCys Ser Gln Pro 100 105 110 Cys Pro Trp Pro Met Ile Glu Lys Leu Pro CysAla Ala Leu Thr Asp 115 120 125 Arg Glu Cys Thr Cys Pro Pro Gly Met PheGln Ser Asn Ala Thr Cys 130 135 140 Ala Pro His Thr Val Cys Pro Val GlyTrp Gly Val Arg Lys Lys Gly 145 150 155 160 Thr Glu Thr Glu Asp Val ArgCys Lys Gln Cys Ala Arg Gly Thr Phe 165 170 175 Ser Asp Val Pro Ser SerVal Met Lys Cys Lys Ala Tyr Thr Asp Cys 180 185 190 Leu Ser Gln Asn LeuVal Val Ile Lys Pro Gly Thr Lys Glu Thr Asp 195 200 205 Asn Val Cys GlyThr Leu Pro Ser Phe Ser Ser Ser Thr Ser Pro Ser 210 215 220 Pro Gly ThrAla Ile Phe Pro Arg Pro Glu His Met Glu Thr His Glu 225 230 235 240 ValPro Ser Ser Thr Tyr Val Pro Lys Gly Met Asn Ser Thr Glu Ser 245 250 255Asn Ser Ser Ala Ser Val Arg Pro Lys Val Leu Ser Ser Ile Gln Glu 260 265270 Gly Thr Val Pro Asp Asn Thr Ser Ser Ala Arg Gly Lys Glu Asp Val 275280 285 Asn Lys Thr Leu Pro Asn Leu Gln Val Val Asn His Gln Gln Gly Pro290 295 300 His His Arg His Ile Leu Lys Leu Leu Pro Ser Met Glu Ala ThrGly 305 310 315 320 Gly Glu Lys Ser Ser Thr Pro Ile Lys Gly Pro Lys ArgGly His Pro 325 330 335 Arg Gln Asn Leu His Lys His Phe Asp Ile Asn GluHis Leu Pro Trp 340 345 350 Met Ile Val Leu Phe Leu Leu Leu Val Leu ValVal Ile Val Val Cys 355 360 365 Ser Ile Arg Lys Ser Ser Arg Thr Leu LysLys Gly Pro Arg Gln Asp 370 375 380 Pro Ser Ala Ile Val Glu Lys Ala GlyLeu Lys Lys Ser Met Thr Pro 385 390 395 400 Thr Gln Asn Arg Glu Lys TrpIle Tyr Tyr Cys Asn Gly His Gly Ile 405 410 415 Asp Ile Leu Lys Leu ValAla Ala Gln Val Gly Ser Gln Trp Lys Asp 420 425 430 Ile Tyr Gln Phe LeuCys Asn Ala Ser Glu Arg Glu Val Ala Ala Phe 435 440 445 Ser Asn Gly TyrThr Ala Asp His Glu Arg Ala Tyr Ala Ala Leu Gln 450 455 460 His Trp ThrIle Arg Gly Pro Glu Ala Ser Leu Ala Gln Leu Ile Ser 465 470 475 480 AlaLeu Arg Gln His Arg Arg Asn Asp Val Val Glu Lys Ile Arg Gly 485 490 495Leu Met Glu Asp Thr Thr Gln Leu Glu Thr Asp Lys Leu Ala Leu Pro 500 505510 Met Ser Pro Ser Pro Leu Ser Pro Ser Pro Ile Pro Ser Pro Asn Ala 515520 525 Lys Leu Glu Asn Ser Ala Leu Leu Thr Val Glu Pro Ser Pro Gln Asp530 535 540 Lys Asn Lys Gly Phe Phe Val Asp Glu Ser Glu Pro Leu Leu ArgCys 545 550 555 560 Asp Ser Thr Ser Ser Gly Ser Ser Ala Leu Ser Arg AsnGly Ser Phe 565 570 575 Ile Thr Lys Glu Lys Lys Asp Thr Val Leu Arg GlnVal Arg Leu Asp 580 585 590 Pro Cys Asp Leu Gln Pro Ile Phe Asp Asp MetLeu His Phe Leu Asn 595 600 605 Pro Glu Glu Leu Arg Val Ile Glu Glu IlePro Gln Ala Glu Asp Lys 610 615 620 Leu Asp Arg Leu Phe Glu Ile Ile GlyVal Lys Ser Gln Glu Ala Ser 625 630 635 640 Gln Thr Leu Leu Asp Ser ValTyr Ser His Leu Pro Asp Leu Leu 645 650 655 3 584 DNA HOMO SAPIENSUNSURE (4)(10)(11)(13)(15)(38) OTHER INFORMATION n= a, g, c or t 3gcgnccgcgn ngngngcaag gtgctgagcg cccctagngc ctcccttgcc gcctccctcc 60tctgcccggc cgtagcagtg cacatggggt gttggaggta gatgggctcc cggccgggag 120gcggcggtgg atgcggcgct gggcagaagc agccgccgat tccagctgcc ccgcgcgccc 180cggccacctt gcgagtcccc ggttcagcca tggggacctc tccgagcagc agcaccgccc 240tcgcctcctg cagccgcatc gcccgccgag ccacagccac gatgatcgcg ggctcccttc 300tcctgcttgg attccttagc accaccacag ctcagccaga acagaaggcc tcgaatctca 360ttggcacata ccgccatgtt gaccgtgcca ccggccaggt gctaacctgt gacaagtgtc 420cagcaggaac ctatgtctct gagcattgta ccaacacaag cctgcgcgtc tgcagcagtt 480gccctgtggg gacctttacc aggcatgaga atggcataga gaaatgccat gactgtagtc 540agccatgccc atggccaatg attgagaaat taccttgtgc tgcc 584 4 125 PRT HOMOSAPIENS 4 Met Gly Thr Ser Pro Ser Ser Ser Thr Ala Leu Ala Ser Cys SerArg 1 5 10 15 Ile Ala Arg Arg Ala Thr Ala Thr Met Ile Ala Gly Ser LeuLeu Leu 20 25 30 Leu Gly Phe Leu Ser Thr Thr Thr Ala Gln Pro Glu Gln LysAla Ser 35 40 45 Asn Leu Ile Gly Thr Tyr Arg His Val Asp Arg Ala Thr GlyGln Val 50 55 60 Leu Thr Cys Asp Lys Cys Pro Ala Gly Thr Tyr Val Ser GluHis Cys 65 70 75 80 Thr Asn Thr Ser Leu Arg Val Cys Ser Ser Cys Pro ValGly Thr Phe 85 90 95 Thr Arg His Glu Asn Gly Ile Glu Lys Cys His Asp CysSer Gln Pro 100 105 110 Cys Pro Trp Pro Met Ile Glu Lys Leu Pro Cys AlaAla 115 120 125 5 588 DNA HOMO SAPIENS UNSURE (4)(10)(11)(13)(15)(38)OTHER INFORMATION n= a, g, c or t 5 gcgnccgcgn ngngngcaag gtgctgagcgcccctagngc ctcccttgcc gcctccctcc 60 tctgcccggc cgtagcagtg cacatggggtgttggaggta gatgggctcc cggccgggag 120 gcggcggtgg atgcggcgct gggcagaagcagccgccgat tccagctgcc ccgcgcgccc 180 cggccacctt gcgagtcccc ggttcagccatggggacctc tccgagcagc agcaccgccc 240 tcggcctcct gcaaccgcat cgcccgccgagccacagcca cgatgatcgc gggctccctt 300 ctcctgcttg gattccttag caccaccacagctcagccag aacagaaggc ctcgaatctc 360 attggcacat accgccatgt tgaccgtgccaccggccagg tgctaacctg tgacaagtgt 420 ccagcaggaa cctatgtctc tgagcattgtaccaacacaa gcctgcgcgt ctgtcagcag 480 tgccctgtgg ggacctttac caggcatgagaatggcatag agaaatgcca tgactgtagt 540 cagccatgcc catggccaat gattgagaaattaccttgtg cctctgcc 588 6 102 PRT HOMO SAPIENS 6 Met Ile Ala Gly Ser LeuLeu Leu Leu Gly Phe Leu Ser Thr Thr Thr 1 5 10 15 Ala Gln Pro Glu GlnLys Ala Ser Asn Leu Ile Gly Thr Tyr Arg His 20 25 30 Val Asp Arg Ala ThrGly Gln Val Leu Thr Cys Asp Lys Cys Pro Ala 35 40 45 Gly Thr Tyr Val SerGlu His Cys Thr Asn Thr Ser Leu Arg Val Cys 50 55 60 Gln Gln Cys Pro ValGly Thr Phe Thr Arg His Glu Asn Gly Ile Glu 65 70 75 80 Lys Cys His AspCys Ser Gln Pro Cys Pro Trp Pro Met Ile Glu Lys 85 90 95 Leu Pro Cys AlaSer Ala 100

What is claimed is:
 1. An isolated TR7 polypeptide comprising the aminoacid sequence of SEQ ID NO:2.
 2. The isolated polypeptide of claim 1consisting of the amino acid sequence of SEQ ID NO:2.
 3. A compositioncomprising an isolated TR7 polypeptide, the isolated TR7 polypeptidecomprising the amino acid sequence of SEQ ID NO:2.
 4. A composition ofclaim 3 wherein the isolated TR7 polypeptide consists of the amino acidsequence of SEQ ID NO:2.
 5. An isolated TR7 polypeptide comprising theamino acid sequence of SEQ ID NO:4.
 6. The isolated TR7 polypeptide ofclaim 5 consisting of the amino acid sequence of SEQ ID NO:4.
 7. Acomposition comprising an isolated TR7 polypeptide, the isolated TR7polypeptide comprising the amino acid sequence of SEQ ID NO:4.
 8. Thecomposition of claim 7 wherein the isolated TR7 polypeptide consists ofthe amino acid sequence of SEQ ID NO:4.
 9. An isolated TR7 polypeptidecomprising at least 20 contiguous amino acids of the amino acid sequenceof SEQ ID NO:6.
 10. The isolated TR7 polypeptide of claim 9 wherein thepolypeptide has at least one cysteine-rich motif characteristic of anextracellular domain of Tumor Necrosis Factor Receptors.
 11. Theisolated polypeptide of claim 9 wherein the polypeptide has at least oneCys-x-x-Cys amino acid motif.
 12. The isolated TR7 polypeptide of claim9 wherein said at least 20 contiguous amino acids of the amino acidsequence of SEQ ID NO:6 are also present in SEQ ID NO:2.
 13. Theisolated TR7 polypeptide of claim 9 wherein said at least 20 contiguousamino acids of the amino acid sequence of SEQ ID NO:6 are also presentin SEQ ID NO:4.
 14. A composition comprising an isolated TR7polypeptide, the isolated TR7 polypeptide comprising at least 20contiguous amino acids of the amino acid sequence of SEQ ID NO:6,wherein the polypeptide has at least one cysteine-rich motifcharacteristic of an extracellular domain of Tumor Necrosis FactorReceptors.
 15. The composition of claim 14 comprising the isolated TR7polypeptide wherein said at least 20 contiguous amino acids of the aminoacid sequence of SEQ ID NO:6 are also present in SEQ ID NO:2.
 16. Thecomposition of claim 14 comprising the isolated TR7 polypeptide whereinsaid at least 20 contiguous amino acids of the amino acid sequence ofSEQ ID NO:6 are also present in SEQ ID NO:4.
 17. The isolated TR7polypeptide of claim 9 which comprises the amino acid sequence of SEQ IDNO:6.